pRF-HU

01/08/10

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Restriction map of pRF-HU

pRF-HU vector sequence in GenBank format

 

LOCUS pRF-HU. 6336 bp DNA circular 19-JAN-2009
DEFINITION Complementary copy of pRF-HU c.
SOURCE
ORGANISM
COMMENT The vector can be used for ectopic integration of any DNA fragment into the
genome of filamentous fungi.

The vector has been designed for the directional cloning of a single PCR
amplicon, utilizing the USER friendly cloning technology from NEB.

Add the following 5' overhangs to the primers for amplification of the two gene
specific homologous recombination flanks:

Gene-U1 (LB forward) 5- GGGTTTAAU - Forward primer sequence
Gene-U2 (LB reverse) 5'- GGACTTAAU - Reverse primer sequence

Digestion of vector prior to cloning:
1. Digest over night with PacI
2. Digest for 2 hours with Nt.Bbv.C IB (nicking enzyme)

Transformation of the vector into the fungi:
The vector is originate designed for Agrobacterium tumefaciens mediated
transformation, but can also be used for protoplast transformation (bipartial
transformation strategies).

COMMENT This file is created by Vector NTI
http://www.invitrogen.com/
COMMENT ORIGDB|EMBL
COMMENT VNTDATE|507680032|
COMMENT VNTDBDATE|507680032|
COMMENT LSOWNER|
COMMENT VNTNAME|pRF-HU.|
COMMENT VNTAUTHORNAME|Rasmus J.N. Frandsen|
COMMENT VNTAUTHOREML|raf@kvl.dk|
COMMENT VNTREPLTYPE|Plasmid
COMMENT VNTEXTCHREPL|Bacteria
FEATURES Location/Qualifiers
rep_origin complement(2679..3296)
/vntifkey="33"
/label=OriV
primer_bind 4915..4936
/vntifkey="28"
/label=Hyg588L
primer_bind complement(5480..5502)
/vntifkey="28"
/label=Hyg588U
primer_bind 5914..5937
/vntifkey="28"
/label=RF-1
misc_feature complement(6046..6071)
/vntifkey="21"
/label=USER
misc_feature complement(3684..3707)
/vntifkey="21"
/label=RF-2
misc_feature complement(3490..3509)
/vntifkey="21"
/label=RF-4
misc_structure complement(1..22)
/vntifkey="88"
/label=LB
CDS complement(149..1298)
/vntifkey="4"
/label=TrfA
CDS complement(1597..2392)
/vntifkey="4"
/label=KanR
CDS complement(4614..5659)
/vntifkey="4"
/label=hph
terminator complement(3894..4610)
/vntifkey="43"
/label=TtrpC
misc_structure complement(3444..3466)
/vntifkey="88"
/label=RB
promoter complement(5660..6024)
/vntifkey="29"
/label=pTrpC
BASE COUNT 1427 a 1796 c 1616 g 1497 t
ORIGIN
1 tttacaccac aatatatcct gccaccagcc agccaacagc tccccgaccg gcagctcggc
61 acaaaatcac cactcgatac aggcagccca tcagtccggg acggcgtcag cgggagagca
121 gatccagccg gaactgaccc cacaaggccc tagcgtttgc aatgcaccag gtcatcattg
181 acccaggcgt gttccaccag gccgctgcct cgcaactctt cgcaggcttc gccgacctgc
241 tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc
301 ttgagcgggt acggctcccg gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc
361 gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg
421 acgatttcct cgtcgatcag gacctggcaa cgggacgttt tcttgccacg gtccaggacg
481 cggaagcggt gcagcagcga caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc
541 atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct tcgtgtaata ccggccattg
601 atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata
661 ggggtgcgct tcgcgtactc caacacctgc tgccacacca gttcgtcatc gtcggcccgc
721 agctcgacgc cggtgtaggt gatcttcacg tccttgttga cgtggaaaat gaccttgttt
781 tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga acagggcaga gcgggccgtg
841 tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc
901 atttccttga tctgctgctt cgtgtgtttc agcaacgcgg cctgcttggc ctcgctgacc
961 tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg
1021 tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt cgagacgacg cgaacgctcc
1081 acggcggccg atggcgcggg cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc
1141 ttggccgtag cttgctggac catcgagccg acggactgga aggtttcgcg gggcgcacgc
1201 atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt
1261 tcttgcctgt atgccttccg gtcaaacgtc cgattcattc accctccttg cgggattgcc
1321 ccgactcacg ccggggcaat gtgcccttat tcctgatttg acccgcctgg tgccttggtg
1381 tccagataat ccaccttatc ggcaatgaag tcggtcccgt agaccgtctg gccgtccttc
1441 tcgtacttgg tattccgaat cttgccctgc acgaatacca gcgacccctt gcccaaatac
1501 ttgccgtggg cctcggcctg agagccaaaa cacttgatgc ggaagaagtc ggtgcgctcc
1561 tgcttgtcgc cggcatcgtt gcgccacatc taggtactaa aacaattcat ccagtaaaat
1621 ataatatttt attttctccc aatcaggctt gatccccagt aagtcaaaaa atagctcgac
1681 atactgttct tccccgatat cctccctgat cgaccggacg cagaaggcaa tgtcatacca
1741 cttgtccgcc ctgccgcttc tcccaagatc aataaagcca cttactttgc catctttcac
1801 aaagatgttg ctgtctccca ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc
1861 cgtctttaaa aaatcataca gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt
1921 ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa tccaattcgg ctaagcggct
1981 gtctaagcta ttcgtatagg gacaatccga tatgtcgatg gagtgaaaga gcctgatgca
2041 ctccgcatac agctcgataa tcttttcagg gctttgttca tcttcatact cttccgagca
2101 aaggacgcca tcggcctcac tcatgagcag attgctccag ccatcatgcc gttcaaagtg
2161 caggaccttt ggaacaggca gctttccttc cagccatagc atcatgtcct tttcccgttc
2221 cacatcatag gtggtccctt tataccggct gtccgtcatt tttaaatata ggttttcatt
2281 ttctcccacc agcttatata ccttagcagg agacattcct tccgtatctt ttacgcagcg
2341 gtatttttcg atcagttttt tcaattccgg tgatattctc attttagcca tttattattt
2401 ccttcctctt ttctacagta tttaaagata ccccaagaag ctaattataa caagacgaac
2461 tccaattcac tgttccttgc attctaaaac cttaaatacc agaaaacagc tttttcaaag
2521 ttgttttcaa agttggcgta taacatagta tcgacggagc cgattttgaa accacaatta
2581 tgggtgatgc tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg
2641 gcgtacgcct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg
2701 ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag
2761 gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg
2821 gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag
2881 gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct
2941 gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca
3001 catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca
3061 gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga
3121 gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc
3181 gaggtatcca caacgccggc ggccgcggtg tctcgcacac ggcttcgacg gcgtttctgg
3241 cgcgtttgca gggccataga cggccgccag cccagcggcg agggcaacca gcccggtgag
3301 cgtcgcaaag gcgctcggtc ttgccttgct cgtcggatct ggaaccctgt ggttggcatg
3361 aacaaacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc cgattattct
3421 aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac actgatagtt
3481 taaactgaag gcgggaaacg acaatctgat catgagcgga gaattaaggg agtcacgaag
3541 ctaaatccgt ttaaacggcg cgccggccgg ccgcgatcgc gcactagtcg ggggatcctc
3601 tagatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
3661 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
3721 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
3781 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
3841 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcctct agaaagaagg
3901 attacctcta aacaagtgta cctgtgcatt ctgggtaaac gactcatagg agagttgtaa
3961 aaaagtttcg gccggcgtat tgggtgttac ggagcattca ctaggcaacc atggttacta
4021 ttgtataccc atcttagtag gaatgatttt cgaggtttat acctacgatg aatgtgtgtc
4081 ctgtaggctt gagagttcaa ggaagaaaca gtgcaattat ctttgcgaac ccaggggctg
4141 gtgacggaat tttcatagtc aagctatcag agtaaagaag aggagcatgt caaagtacaa
4201 ttagagacaa atatatagtc gcgtggagcc aagagcggat tcctcagtct cgtaggtctc
4261 ttgacgaccg ttgatctgct tgatctcgtc tcccgaaaat gaaaatagac tctgctaagc
4321 tattcttctg cttcgccgga gcctgaaggg cgtactaggg ttgcgaggtc caatgcatta
4381 atgcattgca gatgagctgt atctggaaga ggtaaacccg aaacgcgttt tattcttgtt
4441 gacatggagc tattaaatca ctagaaggca ctctttgctg cttggacaaa tgaacgtatc
4501 ttatcgagat cctgaacacc atttgtctca actccggagc tgacatcgac accaacgatc
4561 ttatatccag attcgtcaag ctgtttgatg atttcagtaa cgttaagtgg atcccggtcg
4621 gcatctactc tattcctttg ccctcggacg agtgctgggg cgtcggtttc cactatcggc
4681 gagtacttct acacagccat cggtccagac ggccgcgctt ctgcgggcga tttgtgtacg
4741 cccgacagtc ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc
4801 atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa tgcggagcat
4861 atacgcccgg agccgcggcg atcctgcaag ctccggatgc ctccgctcga agtagcgcgt
4921 ctgctgctcc atacaagcca accacggcct ccagaagaag atgttggcga cctcgtattg
4981 ggaatccccg aacatcgcct cgctccagtc aatgaccgct gttatgcggc cattgtccgt
5041 caggacattg ttggagccga aatccgcgtg cacgaggtgc cggacttcgg ggcagtcctc
5101 ggcccaaagc atcagctcat cgagagcctg cgcgacggac gcactgacgg tgtcgtccat
5161 cacagtttgc cagtgataca catggggatc agcaatcgcg catatgaaat cacgccatgt
5221 agtgtattga ccgattcctt gcggtccgaa tgggccgaac ccgctcgtct ggctaagatc
5281 ggccgcagcg atcgcatcca tggcctccgc gaccggctgc agaacagcgg gcagttcggt
5341 ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg gagatgcaat aggtcaggct
5401 ctcgctgaat tccccaatgt caagcacttc cggaatcggg agcgcggccg atgcaaagtg
5461 ccgataaaca taacgatctt tgtagaaacc atcggcgcag ctatttaccc gcaggacata
5521 tccacgccct cctacatcga agctgaaagc acgagattct tcgccctccg agagctgcat
5581 caggtcggag acgctgtcga acttttcgat cagaaacttc tcgacagacg tcgcggtgag
5641 ttcaggcttt ttcatatcga tgcttgggta gaataggtaa gtcagattga atctgaaata
5701 aagggaggaa gggcgaactt aagaaggtat gaccgggtcg ttcacttacc ttgcttgaca
5761 aacgcaccaa gttatcgtgc accaagcagc agatgataat aatgtcctcg ttcctgtctg
5821 ctaataagag tcacacttcg agcgccgccg ctactgctac aagtggggct gatctgacca
5881 gttgcctaaa tgaaccatct tgtcaaacga cacaaatttt gtgctcaccg cctggacgac
5941 taaaccaaaa taggcattca ttgttgacct ccactagctc cagccaagcc caaaaagtgc
6001 tccttcaata tcatcttctg tcgaggcctg atcatcgatg ggcccgctga ggacttaatt
6061 aaaccctcag cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
6121 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
6181 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
6241 cgccgctgtt gcccgtctca ctggtgaaaa gaaaaaccac cccagtacat taaaaacgtc
6301 cgcaatgtgt tattaagttg tctaagcgtc aatttg
//
                     

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Dette sted blev sidst opdateret 01. August 2010